Wnt/2-catenin signaling has come to the forefront in liver biology. Its role in liver development, regeneration &stem cells is beginning to be understood. We identified its role in liver regeneration &showed activation of the Wnt/2-catenin signaling immediately after partial-hepatectomy (PHx). This was observed as nuclear translocation of 2-catenin protein ensuring G1 to S transition mediated by factors such as Cyclin-D1. In addition, we identified highest 2-catenin expression during early stages of hepatic morphogenesis in liver development &showed that its absence led to compromise in hepatoblast expansion &differentiation into bile ducts &failure of hepatocyte maturation. Recently, we have also identified the role of Wnt/2-catenin in adult liver stem cells or oval cells, where this pathway regulates their emergence &expansion. As we have uncovered several key roles of this pathway, many new questions have arisen! Several of these are of high significance &have taken the form of the current proposal, which is a competing renewal of our previously funded application (1/1/2004-12/31/2008). In the current proposal we want to focus on three aspects of liver biology-development, regeneration &hepatocyte death. We have generated 2-catenin-conditional null mice (KO1) with Foxa3-Cre driven deletion of 2-catenin in hepatoblasts during development. This strategy unveiled the importance of 2-catenin in regulating hepatoblast expansion &differentiation. We propose to identify the molecular basis by which 2-catenin is regulating these two conceptually opposing events during development &hypothesize (based on stem cell paradigm) that differential interaction of 2-catenin occurs temporally with cofactors enabling transactivation of distinct genes that regulate the two processes. We will elucidate the basis of failed biliary differentiation in absence of 2-catenin &examine how lack of 2-catenin retards hepatocyte maturation. Based on the controversy in the role of Wnt/2-catenin signaling in hepatic specification in Zebrafish &Xenopus, we will utilize KO1 to address role of 2-catenin in murine hepatic specification. We have also generated 2-catenin-conditional-null mice (KO2) using Albumin-Cre and identified lack of proliferation in them at 40hrs (peak proliferation in controls) after PHx. We will address the molecular signaling in the absence of 2-catenin that enables a dramatic rescue of hepatocyte proliferation at 72hrs in KO2 mice. While we are beginning to understand the role of canonical Wnt signaling, the role &extent of noncanonical pathways-Wnt/Ca2+ &planar cell polarity pathways;remain obscure &will be investigated in-depth in liver development &regeneration. Finally, based on enhanced apoptosis in hepatocytes lacking 2-catenin, we investigated Fas-&TNF1-mediated injury in the KO2. Interestingly, while KO2 mice were clearly more susceptible to Jo-2 (Fas-ligand) injury than controls, they were resistant to lipopolysaccharide (LPS)-injury. The molecular basis of these findings will be elucidated. Thus, this proposal will be a comprehensive analysis of canonical &noncanonical Wnt signaling in hepatic biology. PUBLIC HEALTH RELEVANCE: Understanding signaling pathways dictating the processes of liver growth, regeneration &development would be critical to identify the molecular basis of many hepatic diseases ranging from developmental anomalies to cancers &hepatic failure due to hepatitis, alcohol &other toxins. Our proposal will comprehensively examine Wnt signaling in liver biology to eventually improve prognosis of liver diseases.